Bridging ELISAs are a type of sandwich ELISA designed for detection of antigens with 2 or more identical binding sites, resulting in the formation of a "bridge" complex (see Fig.1). This format is commonly used for quantification of therapeutic antibodies; additionally, anti-drug antibodies resulting from immune response to a drug can be detected. The bridging ELISA technique is ideally suited for the development of drug antibodies (DA), since it enables pharmacokinetic characterization as well as detection of immunogenicity.
Like all RayBiotech ELISA kits, our bridging sandwich ELISAs are manufactured in Peachtree Corners, GA, backed by a 100% satisfaction guarantee, and are part of a comprehensive menu of competitively priced products and services to advance AD development and discovery.
The drug antibody (DA) bridging ELISAs employ an antibody specific for the drug coated on a 96-well plate. Standards and samples are pipetted into the wells; the drug molecule binds to the immobilized antibody. The wells are washed and the biotinylated anti-drug antibody is then added. After washing away unbound biotinylated antibodies, HRP-conjugated streptavidin is added to the wells, followed by a substrate solution. The intensity of color development in the wells is proportional to the amount of DA bound.
The anti-drug antibody ADA bridging ELISAs employ a DA coated on a 96-well plate. Standards and samples are pipetted into the wells; the anti-drug antibodies present in the sample bind to the immobilized DA. The wells are washed and HRP-conjugated detection antibody is then added, followed by a substrate solution. The intensity of color development in the wells is proportional to the amount of ADA bound.
How It Works for Drug Antibodies (DA)
How It Works for Anti-Drug Antibodies (ADA)
- Reliable, robust sandwich ELISA format
- Assays available for many FDA-approved biologics
- Compatible with serum, plasma, and many other sample types
- Quantifying DA in a variety of matrices
- Pharmacokinetic analysis
- Immunogenicity characterization
- Detection of unwanted immune responses to biotherapeutics
- Custom kits available (contact us for a quote)
Contents of Kit
- 96-well pre-coated strip plate (12x8)
- Wash Buffer Concentrate (20x)
- DA or ADA Standard
- Sample Diluent Buffers
- Labeled Detection Antibody
- HRP-Streptavidin concentrate*
- TMB One-Step Substrate Reagent
- Stop Solution